Biofabricated Humanized Insulin Producing Neo-Organs Generates Secondary Neo-Organoids Through Ectopic Transplantation
Developing transplantable humanized endocrine grafts for long-term management of disorders like uncontrolled diabetes mellitus has been one of the most challenging concerns in tissue and organ bioengineering. Hence, we aimed to develop bio/immune compatible and transplantable humanized insulin producing neo-organs through biofabrication of rat spleen which acts as secondary neo-organoid after ectopic transplantation.
An efficient process was developed using splenic artery perfusion to generate completely cell deficient decellularized splenic matrices (DSM). Microscopic, vascular tree imaging, mechanical strength and other required analysis were performed before and after decellularization. Biofabrication of humanized insulin producing neo-organ was performed through infusion of hyperglycaemic challenged trans-differentiated human hepatic progenitor cells (hHPCs) in DSM. Intra-peritoneal transplantation of biofabricated insulin producing humanized neo-organs in rats was done to evaluate the bio/immune compatibility.
The biofabrication strategy generated highly intact DSM with preserved organ architecture, extracellular matrix and organ vasculature. Repopulation of DSM with glucose responsive cells showed enhanced engraftment efficiency, survival and proliferation. Bioengineered humanized insulin producing neo-organs showed higher amount of insulin secretion than conventional 2D-cultured cells in response to hyperglycaemic stimuli ex vivo. Glucose responsive humanized neo-organs survived in vivo without eliciting immunological or fibrotic responses and proved to be secondary neo-organoids at ectopic site with the development of neo-vasculature and responded to hyperglycaemic challenge.
The present study provides first proof-of-concept for biofabricating bio/immune compatible, humanized insulin producing neo-organoids post-ectopic transplantation which provides more suitable functional biological implants for long-term management of uncontrolled diabetes mellitus.
KeywordsBioengineering Endocrine neo-organ Diabetes management Insulin secretion Biocompatibility Decellularization Stem cells
All authors thank to CLRD, DCMS for providing necessary infrastructure to conduct the study. We thank to RUSKA Lab, Acharya NG Ranga Agricultural University, Rajendra Nagar, Hyderabad for helping us to perform SEM studies.
SKV and AAK conceptualized and designed the study. SKV, AB, CL, and RN performed the experiments. SABP and MAH gave inputs, wrote the clinical part of the study and provided required infrastructure. SKV performed statistical analysis of the data and designed the images. SKV, AB, CL and RN wrote and formatted the manuscript. Proof-reading of the manuscript was done by SKV, AB, AAK, SABP and MAH. Language editing was done by SKV and AAK.
Conflict of interest
Sandeep Kumar Vishwakarma, Chandrakala Lakkireddy, Avinash Bardia, Raju Nagarapu, Syed Ameer Basha Paspala, Md. Aejaz Habeeb, and Aleem Ahmed Khan declare that they have no conflict of interest.
The study design was approved by the Institutional Review Board of Deccan College of Medical Sciences, Hyderabad, Telangana, India. All the animal experiments were performed according to the guidelines of Institutional Animal Ethics Committee of Deccan College of Medical Sciences, Hyderabad, Telangana, India.
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